Q: Does this antibody cross-react with GFP epitopes? As I would like to use both GFP and mCherry antibodies during histochemistry I would not like them to cross-react.

A: mCherry and GFP share just 29% sequence similarity, so this antibody is not predicted to cross-react to GFP and has never shown any ability to detect GFP in testing.

mCherry Antibody

Novus Biologicals | Catalog # NBP3-50195

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Product Specifications
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species Reactivity

Non-species specific

Applications

Western Blot, ELISA

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

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Product Specifications

Immunogen

The immunogen is a mCherry mutant variant fusion protein of RFP corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.

Specificity

Expect reactivity against mCherry, RFP and its variants: tdTomato, mBanana, mOrange, mPlum, mOrange and mStrawberry. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum and purified and partially purified mCherry. No reaction was observed against Human, Mouse or Rat serum proteins. ELISA was used to confirm specificity at less than 0.1% of target signal.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Applications for mCherry Antibody

Application

Recommended Usage

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.

Application Notes

This antibody is designed to detect mCherry, RFP, and its variants, & has been tested by ELISA and Western blot and is intended for use in immunological assays including ELISA, western blotting, immunofluorescence, and fluorescence activated cell sorting (FACS). Researchers should determine optimal titers for applications that are not stated. In addition, we performed conjugation of RFP antibodies to either fluorescent dyes, biotin or horseradish peroxidase to further facilitate RFP protein detection and quantification.

Suggested applications: IF; IHC; IP; Multiplex

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: mCherry

mCherry is a monomeric red fluorescent protein (mRFP) belonging to the mFruits family which is brighter and more photostable compared to the first-generation mRFP1, making them ideal for fluorescence microscopy (1). mCherry has an excitation maximum at 587 nm and an emission maximum at 610 nm. mCherry protein was derived from DsRed, a red fluorescent protein from the coral Discosoma (disc anemone) (2). The red chromophore of DsRed has a similar topology to GFP, the green fluorescent protein isolated from the jellyfish Aequorea Victoria, but has extended pi-electron conjugation resulting in red-shifted absorbance and emission (3). mCherry is 236 amino acids (aa) in length with a theoretical molecular weight of 28 kDa and has a crystal structure with the chromophore forming a central helix shielded within an eleven-stranded beta-barrel (3).

mCherry can be used as a long-wavelength hetero-FRET (fluorescence resonance energy transfer) acceptor and probe for homoFRET experiments given its high peak molar absorptivity, folding efficiency, and superior spectral properties (4). Additionally, because mCherry does not interfere with other plasmids or alter the growth of Legionella species during intracellular growth, it can be used for constitutive gene expression in a variety of gram-negative bacterial species (5). For example, a plasmid developed to constitutively express mCherry under the Ptac promoter has been used in several Legionella species including L. pneumophila, the causative agent of Legionnaires' disease (5).

References

1. Shaner, N. C., Steinbach, P. A., & Tsien, R. Y. (2005). A guide to choosing fluorescent proteins. Nature Methods, 2(12), 905-909. doi:10.1038/nmeth819

2. Bevis, B. J., & Glick, B. S. (2002). Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed). Nature Biotechnology, 20(1), 83-87. https://doi.org/10.1038/nbt0102-83

3. Wall, M. A., Socolich, M., & Ranganathan, R. (2000). The structural basis for red fluorescence in the tetrameric GFP homolog DsRed. Nature Structural Biology, 7(12), 1133-1138. https://doi.org/10.1038/81992

4. Akrap, N., Seidel, T., & Barisas, B. G. (2010). Forster distances for fluorescence resonant energy transfer between mCherry and other visible fluorescent proteins. Analytical Biochemistry, 402(1), 105-106. https://doi.org/10.1016/j.ab.2010.03.026

5. Gebhardt, M. J., Jacobson, R. K., & Shuman, H. A. (2017). Seeing red; the development of pON.mCherry, a broad-host range constitutive expression plasmid for Gram-negative bacteria. Plos One, 12(3), e0173116. https://doi.org/10.1371/journal.pone.0173116

Long Name

mCherry

Alternate Names

DSRED, red fluorescent protein mCherry, Red Fluoroscent Protein

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Product Documents for mCherry Antibody

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Certificate of Analysis

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Product Specific Notices for mCherry Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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